Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: Comparative drug response profiles of 2D cell cultures and MCSs. ( a – c ) Drug response curves for LK0917, LK0902, and LK1108 for 2D and MCSs cultures treated with 170–17000 µM of doxorubicin; 3.33–333 µM of cisplatin; and 2.2–220 µM of methotrexate. ( d ) IC 50 (µM) values calculated from the drug response curves for both 2D and MCSs of all cell types for all drugs. Significantly large differences between the IC 50 values of 2D and MCSs are denoted with “ # ”. Each IC 50 value is the average of three independent experiments (n = 3) with triplicates set for each concentration.

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: Real-time monitoring of calcein-AM uptake in 2D and MCSs. ( a ) Elucidate calcein-AM uptake in 2D cell cultures and MCSs obtained from LK0917- MCS 17 (upper panel), LK0902- MCS 02 (middle panel), and LK1108- MCS 08 (lower panel) and over a time span of 10 hours with image acquisition at 20-minute intervals (scale bar, 200 μm). ( b ) Heat map pseudo color images of MCSs for differential calcein-AM uptake for the three cell lines (scale bar, 200 μm, scale bar on the right represents the pixel intensity of green fluorescence). ( c ) Mean fluorescence intensity profile with respect to time (12 hours) in the 2D cell cultures (LK0917, LK0902, and LK1108) and MCSs (MCS 17 , MCS 02 , and MCS 08 ) respectively. ( d ) Total accumulated calcein over time (12 hours) in 2D and MCSs for all three cell lines. ( e ) Total accumulated calcein profiles of MCSs obtained from different cell densities. The data are shown as mean ± SD; ***p < 0.001, **p < 0.05, *p = 0.019, and ns = non-significant (n = 3).

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Fluorescence

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: ROS activity of all cell lines in 2D and MCSs. ( a ) Live-cell fluorescence images of the 2D (LK0917, LK0902, and LK1108) and MCSs (MCS 17 , MCS 02 , and MCS 08 ) cultures obtained from cell lines in the presence of DCFDA over a period of 60 minutes with image acquisition at 20-minute intervals (upper, middle and lower panel respectively), scale bar, 200 μm). ( b ) Redox state in 2D and MCSs cultures. Data are presented as the mean ± SD; ***p < 0.001, **p < 0.05, *p = 0.019, and no significant differences were observed in case of 2D (n = 3). ( c ) Heat map pseudo color images of MCSs for differential redox status for the three cell lines (scale bar, 200 μm).

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Activity Assay, Fluorescence

Journal: Scientific Reports

Article Title: Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids

doi: 10.1038/s41598-019-56273-6

Figure Lengend Snippet: ( a –c) Median fluorescence intensity (MFI) values for 2D and MCSs cultures of LK0917, LK0902, and LK1108 in presence of different ABC pump inhibitors for MDR1, MRP1, and BCRP. ( d ) MFI for MDR1 pump inhibitor (verapamil) and its corresponding live cell imaging for 2D and MCSs cultures. ( e ) Total calcein-AM uptake for the entire course of 10 hours +/− verapamil for 2D and MCSs. The data are shown as a mean of ± SD, ***p < 0.001, **p < 0.05. ( f ) Elucidate calcein-AM uptake in 2D for LK0917, LK0902 and LK1108 in presence and absence of verapamil over a time span of 10 hours with images taken after every 20 minutes (images shown here are at 2 h interval). Scale bar 200 μm.

Article Snippet: For the generation of MCSs sized 300–500 μm, 200 μL of LK0917 (MCS 17 ), LK0902 (MCS 02 ), and LK1108 (MCS 08 ) single cell suspensions were seeded in ultra-low attachment (ULA) plates (Corning Life Sciences, Massachusetts, USA) at varying cell densities in the range of 0.25–0.75 × 10 5 cells/mL.

Techniques: Fluorescence, Live Cell Imaging

Journal: EJHaem

Article Title: Altered type 1 interferon responses in alloimmunized and nonalloimmunized patients with sickle cell disease

doi: 10.1002/jha2.270

Figure Lengend Snippet: Interferon-stimulated genes in blood leukocytes of SS patients compared to AA controls. (A) Relative expression of interferon-stimulated genes (MxA, Ly6E, IFIT3, IFI44, IFI44L, ISG15, and IFI27) by qPCR using RNA from whole blood of SS patients and AA controls. (B) Equation used to calculate IFN α / β gene scores. (C) IFN α / β gene scores calculated from expression of ISGs in (A). (D) IFN α / β gene scores of AA controls, alloimmunized, and nonalloimmunized SS patients. * p < 0.05, ** p < 0.01

Article Snippet: Samples were thawed, and single-cell suspensions of blood leukocytes were obtained using Thaw-Lyse buffer (Smart Tube, Inc) according to manufacturer’s instructions.

Techniques: Expressing

Journal: EJHaem

Article Title: Altered type 1 interferon responses in alloimmunized and nonalloimmunized patients with sickle cell disease

doi: 10.1002/jha2.270

Figure Lengend Snippet: Reduced IFN β -induced expression of ISGs by monocytes from whole blood of patients with SCD. Peripheral blood leukocytes from AA controls and patients with SCD (SS) were stimulated in whole blood with or without IFN β . (A) Representative flow cytometric analysis of CD14+CD64+ monocytes, gated on HLADR + nonlymphocytes. Numbers on plots indicate the percentage of cells within the drawn gate. (B) Representative histograms and (C) quantification of Siglec-1 and CD38 expression by unstimulated and IFN β -stimulated CD14 + CD64 + monocytes gated in (A). ** p < 0.01, *** p < 0.001

Article Snippet: Samples were thawed, and single-cell suspensions of blood leukocytes were obtained using Thaw-Lyse buffer (Smart Tube, Inc) according to manufacturer’s instructions.

Techniques: Expressing

Journal: EJHaem

Article Title: Altered type 1 interferon responses in alloimmunized and nonalloimmunized patients with sickle cell disease

doi: 10.1002/jha2.270

Figure Lengend Snippet: Whole blood IFN β stimulation failed to induce CD38 in T cells from patients with SCD. Peripheral blood leukocytes from AA controls and patients with SCD (SS) were stimulated in whole blood with or without IFN β . (A,F) Representative flow cytometric analysis of (A) CD3 + T cells and (F) CD19 + B cells, gated on CD66b − nonneutrophils. Numbers on plots indicate the percentage of cells within the drawn gate. (B,G) Representative histograms and (E,J) quantification of CD38 expression by unstimulated and IFN β -stimulated T and B cells gated in (A) and (F). (C,H) Representative histograms and (D,I) quantification of CD38 expression on AA controls and SS patients with or without alloimmunization. p < 0.05, ** p < 0.01

Article Snippet: Samples were thawed, and single-cell suspensions of blood leukocytes were obtained using Thaw-Lyse buffer (Smart Tube, Inc) according to manufacturer’s instructions.

Techniques: Expressing